LOCUS       Exported               10136 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM410, complete sequence.
ACCESSION   FJ389188
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10136)
  AUTHORS   Chou H-H., Berthet J, Marx CJ.
  TITLE     Growth rate modulates the selective advantage of recurrent 
            transposition mutations arising from experimental populations grown 
            in a metal-limited environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10136)
  AUTHORS   Marx CJ, Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 10136)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10136
                     /organism="Cloning vector pCM410"
                     /mol_type="other DNA"
                     /db_xref="taxon:646102"
     rep_origin      4..712
                     /label=oriV
                     /note="oriV"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1180..1735
                     /label=colE1
                     /note="colE1"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      2011..2053
                     /regulatory_class="promoter"
                     /note="Plac"
     promoter        2017..2047
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2055..2071
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2079..2095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2206..2306
                     /regulatory_class="promoter"
                     /note="PmxaF"
     misc_feature    2307..2574
                     /note="5 upstream region of Methylobacterium extorquens 
                     AM1 moxF"
     primer_bind     2446..2462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2475..2531
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             2622..3749
                     /codon_start=1
                     /gene="flhA"
                     /product="glutathione-dependent formaldehyde dehydrogenase"
                     /label=flhA
                     /note="GD-FALDH"
                     /protein_id="ACR45016.1"
                     /translation="MRTRAAVALEAGKPLEVMEVNLEGPKAGEVMVEIKATGICHTDEF
                     TLSGADPEGLFPSILGHEGAGVVVEVGPGVTSVKPGDHVIPLYTPECRQCASCLSGKTN
                     LCTAIRATQGQGLMPDGTSRFSMLDGTPIFHYMGCSTFSNYTVLPEIAVAKVREDAPFD
                     KICYIGCGVTTGIGAVINTAKVEIGAKAVVFGLGGIGLNVLQGLRLAGADMIIGVDLND
                     DKKPMAEHFGMTHFINPKNCENVVQEIVNLTKTPFDQIGGADYSFDCTGNVKVMRDALE
                     CTHRGWGQSIIIGVAPAGAEISTRPFQLVTGRVWKGTAFGGARGRTDVPQIVDWYMDGK
                     IEIDPMITHTLSLDDINKGFDLMHAGESIRSVVLY"
     gene            2622..3749
                     /gene="flhA"
                     /label=flhA
     CDS             3803..4642
                     /codon_start=1
                     /gene="fghA"
                     /product="S-formylglutathione hydrolase"
                     /label=fghA
                     /note="S"
                     /protein_id="ACR45017.1"
                     /translation="MTLAYETVSENRSFGGIQGVYRHQSQATGTPMTFAIYLPPDARHG
                     KVPVLWYLSGLTCTHENAMTKAGAQEWAAEYGIAVIFPDTSPRGEGVANDETYDLGQGA
                     GFYVDATEAPWAPHFRMWHYVTHELPELVFNNFPLDREAQGITGHSMGGHGALTIAMTF
                     PERYRSVSAFAPIAHPSESDWGRKQFAAYLGDDKAAWKRHDSTILMREKGYPGEVLIDQ
                     GASDQFLDLLKPEALAHAMAERRQPGTFRMQQGYDHSYFFVQSFMADHIRWHAERLG"
     gene            3803..4642
                     /gene="fghA"
                     /label=fghA
     primer_bind     complement(4694..4710)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5131..5796)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor protein"
                     /label=tetR
                     /note="TetR"
                     /protein_id="ACR45018.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGAKKPLLS"
     gene            complement(5131..5796)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(6574..7389)
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aph
                     /note="Aph; kanamycin-resistance protein"
                     /protein_id="ACR45020.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(6574..7389)
                     /gene="aph"
                     /label=aph
     CDS             complement(8061..9209)
                     /codon_start=1
                     /gene="trfA"
                     /product="plasmid replication protein TrfA"
                     /label=trfA
                     /protein_id="ACR45019.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(8061..9209)
                     /gene="trfA"
                     /label=trfA
     oriT            9391..10129
                     /label=origin of transfer
                     /note="origin of transfer"
     CDS             complement(9481..9852)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9885..9994
                     /note="incP origin of transfer"